Effect and mechanism study of different parts of Ningxia Dense Point Hemp Lizard on cisplatin resistant gastric cancer model nude mice
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摘要: 目的 通过建立人胃癌耐药细胞株MKN-45/顺铂皮下瘤裸鼠模型，探讨宁夏密点麻蜥身体不同部位联合顺铂对顺铂耐药胃癌模型裸鼠的干预作用。方法 将36只BALB/c裸鼠均注射MKN-45/顺铂单细胞悬液于右前腋侧皮下制备胃癌裸鼠顺铂耐药模型。造模成功后随机分为模型组、顺铂组、宁夏密点麻蜥头部+顺铂组、宁夏密点麻蜥四肢+顺铂组、宁夏密点麻蜥躯干+顺铂组、宁夏密点麻蜥尾部+顺铂组共6组。除模型组外，余组均给予相应药物治疗28 d。测定各组裸鼠肿瘤体积大小、肿瘤重量及抑瘤率；分别采用免疫印迹法及免疫组织化学法检测各组裸鼠瘤组织中耐药相关蛋白P-糖蛋白(P-gp)、多药耐药蛋白1(MRP1)、B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)表达水平；采用TUNEL法检测各组裸鼠瘤组织中耐药细胞凋亡情况。结果 与模型组比较，各给药组肿瘤重量均减轻(P < 0.05)，各给药组中P-gp、MRP1、Bcl-2的表达均显著降低(P < 0.05)，Bax的表达均显著增高(P < 0.05)，耐药细胞凋亡率均显著增高(P < 0.05)；与顺铂组比较，宁夏密点麻蜥不同部位+顺铂各组肿瘤重量均明显减轻(P < 0.05)，以宁夏密点麻蜥尾部+顺铂组肿瘤重量减轻最显著(P < 0.05)，且抑瘤率最高；P-gp、MRP-1、Bcl-2的表达均降低(P < 0.05)，Bax的表达均升高(P < 0.05)，且前两者均以宁夏密点麻蜥尾部+顺铂组最明显(P < 0.05)；各组裸鼠瘤组织中耐药细胞凋亡率均显著增高(P < 0.05)，以宁夏密点麻蜥尾部+顺铂组凋亡率增高最显著(P < 0.05)。结论 宁夏密点麻蜥身体不同部位联合顺铂均可有不同程度逆转胃癌裸鼠顺铂耐药的作用，以宁夏密点麻蜥尾部+顺铂组增敏提效作用最为明显。Abstract: Objective To establish a human gastric cancer resistant cell line MKN-45/DDP subcutaneous tumor model in nude mice, and to investigate the intervention of different body parts of Ningxia Dense Point Lizard combined with cisplatin on the cisplatin resistant gastric cancer model in nude mice.Methods Thirty-six BALB/c nude mice were injected with MKN-45 /DDP single cell suspension subcutively in the right anterior axillary side to prepare cisplatin resistant models of nude mice with gastric cancer. After successful modeling, they were randomly divided into 6 groups: model group, cisplatin group, head+cisplatin group, all fours+cisplatin group, body+cisplatin group, and afterbody+cisplatin group. Except model group, other groups were treated with corresponding drugs for 28 days. Tumor size, tumor weight and tumor inhibition rate were determined in each group. The expression levels of drug-resistant protein P-glycoprotein (P-gp), multidrug-resistant protein 1 (MRP1), B-cell lymphoma-2 (Bcl-2) and Bcl-2 associated X protein (Bax) in tumor tissue of nude mice in each group were detected by Western blot and immunohistochemistry. TUNEL method was used to detect the apoptosis of drug-resistant cells in tumor tissue of nude mice.Results Compared with model group, tumor weight was decreased in all treatment groups (P < 0.05), the expression of P-gp, MRP1 and Bcl-2 in all treatment groups were significantly decreased (P < 0.05), the expression of Bax was significantly increased (P < 0.05), and the apoptosis rate of drug-resistant cells was significantly increased (P < 0.05). Compared with cisplatin group, the tumor weight of different body parts of Ningxia Dense Point Lizard+cisplatin group was significantly reduced (P < 0.05), and the tumor weight in the afterbody+cisplatin group was the most obvious reduction (P < 0.05), and the tumor inhibitory rate was the highest. The expressions of P-gp, MRP1 and Bcl-2 were decreased (P < 0.05), while the expressions of Bax were increased (P < 0.05), and the first two were most obvious in the afterbody+cisplatin group (P < 0.05). The apoptotic rate of drug-resistant cells in the tumor tissue of nude mice in all groups was significantly increased (P < 0.05), and the apoptotic rate was the most significantly increased in the afterbody+cisplatin group (P < 0.05).Conclusion The combination of cisplatin with different body parts of Ningxia Dense Point Lizard can reverse cisplatin resistance in nude mice with gastric cancer, and the afterbody+cisplatin group has the most obvious sensitization and enhancing effect.
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Key words:
- Ningxia Dense Point Hemp Lizard /
- gastric cancer /
- cisplatin /
- drug resistance
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表 1 各组裸鼠肿瘤重量及抑瘤率比较
X±S 组别 例数 肿瘤重量/g 抑瘤率/% 模型组 6 0.70±0.99 0 顺铂组 6 0.56±0.131) 20 宁夏密点麻蜥头部+顺铂组 6 0.41±0.141)2) 41 宁夏密点麻蜥四肢+顺铂组 6 0.39±0.121)2) 45 宁夏密点麻蜥躯干+顺铂组 6 0.31±0.101)2) 55 宁夏密点麻蜥尾部+顺铂组 6 0.16±0.071)2)3) 77 与模型组比较，1)P < 0.05；与顺铂组比较，2)P < 0.05；各联合组组内比较，3)P < 0.05。 
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表 2 各组裸鼠肿瘤组织中P-gp、MRP1、Bcl-2、Bax的表达情况
X±S 组别 例数 P-gp MRP1 Bcl-2 Bax 模型组 6 0.049±0.007 0.222±0.050 0.060±0.004 0.001±0.000 顺铂组 6 0.038±0.0031) 0.156±0.0051) 0.036±0.0011) 0.007±0.0011) 宁夏密点麻蜥头部+顺铂组 6 0.027±0.0081)2) 0.125±0.0031)2) 0.029±0.0021)2) 0.012±0.0011)2) 宁夏密点麻蜥四肢+顺铂组 6 0.022±0.0041)2) 0.110±0.0031)2) 0.021±0.0061)2) 0.013±0.0021)2) 宁夏密点麻蜥躯干+顺铂组 6 0.011±0.0051)2) 0.081±0.0051)2) 0.018±0.0021)2) 0.016±0.0071)2) 宁夏密点麻蜥尾部+顺铂组 6 0.009±0.0041)2) 0.041±0.0021)2)3) 0.008±0.0011)2)3) 0.036±0.0011)2)3) 与模型组比较，1)P < 0.05；与顺铂组比较，2)P < 0.05；各联合组组内比较，3)P < 0.05。
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表 3 各组裸鼠肿瘤组织中耐药细胞凋亡情况
%, X±S 组别 例数 细胞凋亡率 模型组 6 0.046±0.015 顺铂组 6 0.110±0.0201) 宁夏密点麻蜥头部+顺铂组 6 0.453±0.0251)2) 宁夏密点麻蜥四肢+顺铂组 6 0.530±0.0451)2) 宁夏密点麻蜥躯干+顺铂组 6 0.773±0.0301)2) 宁夏密点麻蜥尾部+顺铂组 6 0.830±0.0361)2)3) 与模型组比较，1)P < 0.05；与顺铂组比较，2)P < 0.05；各联合组组内比较，3)P < 0.05。
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[1] Sung H, Ferlay J, Siegel RL, et al. Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J]. CA Cancer J Clin, 2021, 71(3): 209-249. doi: 10.3322/caac.21660
[2] 艾梦环, 关芳, 王骄, 等. 复方蜥蜴散含药大鼠血清对人胃癌细胞MGC-803增殖、凋亡及周期的影响[J]. 辽宁中医杂志, 2020, 47(1): 179-182, 224. https://www.cnki.com.cn/Article/CJFDTOTAL-LNZY202001059.htm
[3] 关芳, 艾梦环, 王骄, 等. 复方蜥蜴散不同微粒组合剂含药大鼠血清对人SGC-7901胃癌细胞SIRT1、P53的影响[J]. 山西中医, 2019, 35(9): 57-59. https://www.cnki.com.cn/Article/CJFDTOTAL-SHIX201909029.htm
[4] 李卫强, 王骄, 关芳, 等. 宁夏密点麻蜥不同部位含药大鼠血清对人胃癌细胞凋亡的影响研究[J]. 天然产物研究与开发, 2019, 31(8): 1447-1451. https://www.cnki.com.cn/Article/CJFDTOTAL-TRCW201908021.htm
[5] 刘颖慧. α-常春藤皂苷调控人胃癌耐药细胞增殖和凋亡机制研究[D]. 武汉: 武汉大学, 2020.
[6] 赵欢, 吕鹏, 潘一鸣, 等. 复方浙贝颗粒联合顺铂对L1210/CDDP移植瘤模型生存期及多药耐药基因影响的研究[J]. 中国中医急症, 2019, 28(11): 1920-1924. doi: 10.3969/j.issn.1004-745X.2019.11.010
[7] 陈奇. 中药药理研究方法学[M]. 2版. 北京: 人民卫生出版社, 2006: 1169-1169.
[8] 师林, 黄圆圆, 柯斌. 加味龟鹿二仙胶汤对肺癌耐药裸鼠化疗药敏性的改善作用[J]. 中成药, 2021, 43(11): 2976-2981. doi: 10.3969/j.issn.1001-1528.2021.11.008
[9] 王冬博, 聂晶, 武慧娜, 等. 铂类抗肿瘤药物耐药机制的研究进展和应对策略[J]. 药学实践杂志, 2022, 40(4): 302-308. https://www.cnki.com.cn/Article/CJFDTOTAL-YXSJ202204001.htm
[10] 卢明华, 陶功财, 方亚男, 等. 缺氧导致胃癌化疗耐药的研究进展[J]. 中国肿瘤临床, 2019, 46(13): 700-703. doi: 10.3969/j.issn.1000-8179.2019.13.437
[11] 余磊. 8种治疗胃癌常用抗肿瘤药物造成不良反应病例报告研究[J]. 临床医药文献电子杂志, 2019, 6(76): 162. https://www.cnki.com.cn/Article/CJFDTOTAL-LCWX201976127.htm
[12] 朱西杰, 杨利侠, 梁岩. 蜥蜴临证辨析[J]. 山西中医, 2002, 18(4): 45-46. https://www.cnki.com.cn/Article/CJFDTOTAL-SHIX200204034.htm
[13] 《全国中草药汇编》编写组. 全国中草药汇编[M]. 北京: 人民卫生出版社, 1983: 660-661.
[14] 朱西杰, 李卫强, 钱月慧. 中华虫药宁夏蜥蜴[M]. 北京: 中国中医药出版社, 2014: 16-16.
[15] 景燕燕, 白星, 李卫强. 蜥蜴胃康方对胃癌浸润转移的干预作用及机制研究[J]. 时珍国医国药, 2022, 33(4): 783-786. https://www.cnki.com.cn/Article/CJFDTOTAL-SZGY202204004.htm
[16] 陈绍军. 从蜥蜴尾部再生看肿瘤对机体的危害[J]. 医学理论与实践, 2004, 17(1): 4-6. https://www.cnki.com.cn/Article/CJFDTOTAL-YXLL200401002.htm
[17] 刘洪盛, 王裕, 张丹, 等. 青果多糖提取物对人胃癌5-FU耐药细胞的逆转作用[J]. 中国医院药学杂志, 2021, 41(19): 1945-1949. https://www.cnki.com.cn/Article/CJFDTOTAL-ZGYZ202119004.htm
[18] Xue JJ, Liu Y, Yang DR, et al. Dual-phase 99mTc-MIBI imaging and the expressions of P-gp, GST-π, and MRP1 in hyperparathyroidism[J]. Nucl Med Commun, 2017, 38(10): 868-874.
[19] 李蕊. MRP1、GLUT1在骨肉瘤中的表达水平及其与临床病理特征的关系[J]. 中国民康医学, 2020, 32(24): 4-8. https://www.cnki.com.cn/Article/CJFDTOTAL-ZMYX202024002.htm
[20] 高洁, 贾翌江, 阿依江, 等. 肿瘤多药耐药机制及逆转耐药研究进展[J]. 现代肿瘤医学, 2022, 30(21): 3991-3995. https://www.cnki.com.cn/Article/CJFDTOTAL-SXZL202221033.htm
[21] Huang WJ, Ruan S, Wen F, et al. Multidrug resistance of gastric cancer: the mechanisms and Chinese medicine reversal agents[J]. Cancer Manag Res, 2020, 12: 12385-12394.
[22] 孙亚棋, 聂旭阳, 高敬林, 等. P-糖蛋白介导的川芎嗪逆转肿瘤细胞多药耐药机制研究进展[J]. 中国新药杂志, 2022, 31(16): 1580-1583. https://www.cnki.com.cn/Article/CJFDTOTAL-ZXYZ202216005.htm
[23] 刘清华, 张瑞瑞, 刘磊, 等. p70 s6k-siRNA在顺铂耐药胃癌细胞中的作用[J]. 徐州医科大学学报, 2021, 41(7): 480-486. https://www.cnki.com.cn/Article/CJFDTOTAL-XZYX202107004.htm
[24] 王昌高, 昝慧, 张万里, 等. 胡桃醌对胃癌细胞耐药性及细胞中ANXA2、ERCC1表达和AKT/mTOR信号通路的影响[J]. 胃肠病学和肝病学杂志, 2020, 29(2): 136-141.
[25] 陈骢, 郝健淇, 彭皓宁, 等. BCL-2家族凋亡调控作用及其介导的抗肿瘤药物治疗后耐药的研究进展[J]. 中国胸心血管外科临床杂志, 2023, 30(1): 140-148.
[26] 兰炜兰. 基于PI3K/AKT/mTOR信号通路研究八宝丹逆转胃癌多药耐药的作用机制[D]. 福州: 福建中医药大学, 2021.
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